Myasthenia gravis (MG) is an antibody-mediated autoimmune disease and its pathogenesis is closely related to CD4 + T cells. In recent years, gut microbiota is considered to play an important role in the pathogenesis of MG. Astragaloside IV (AS-IV) is one of the main active components extracted from Astragalus membranaceus and has immunomodulatory effects. To study the immunomodulatory effect of AS-IV and the changes of gut microbiota on experimental autoimmune myasthenia gravis (EAMG) mice, we explore the possible mechanism of AS-IV in improving MG.In this study, network pharmacology was utilized to screen the crucial targets of AS-IV in the treatment of MG. Subsequently, a Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis was performed to identify potential pathways through which AS-IV acts against MG. Furthermore, experimental investigations were conducted to validate the underlying mechanism of AS-IV in MG treatment. Before modeling, 5 mice were randomly selected as the control group (CFA group), and the other 10 were induced to EAMG model. These mice were randomly divided into EAMG group and EAMG + AS-IV group, n = 5/group. In EAMG + AS-IV group, AS-IV was administered by gavage. CFA and EAMG groups were given the same volume of PBS. Body weight, grip strength and clinical symptoms were assessed and recorded weekly. At the last administration, the feces were collected for 16S RNA microbiota analysis. The levels of Treg, Th1 and Th17 cells in spleen and Th1 and Th17 cells in thymus were detected by flow cytometry. The levels of IFN-γ, IL-17 and TGF-β in serum were measured by ELISA. Furthermore, fecal microbial transplantation (FMT) experiments were performed for exploring the influence of changed intestinal flora on EAMG. After EAMG model was induced, the mice were treated with antibiotics daily for 4 weeks to germ-free. Then germ-free EAMG mice were randomly divided into two groups: FMT EAMG group, FMT AS-IV group, n = 3/group. Fecal extractions from EAMG and EAMG + AS-IV groups as gathered above were used to administered daily to the respective groups for 4 weeks. Body weight, grip strength and clinical symptoms were assessed and recorded weekly. The levels of Treg, Th1 and Th17 cells in spleen and Th1 and Th17 cells in thymus were detected at the last administration. The levels of IFN-γ, IL-17 and TGF-β in serum were measured by ELISA.The network pharmacology and KEGG pathway analysis revealed that AS-IV regulates T cell pathways, including T cell receptor signaling pathway and Th17 cell differentiation, suggesting its potential in improving MG. Further experimental verification demonstrated that AS-IV administration improved muscle strength and body weight, reduced the level of Th1 and Th17 cells, enhanced the level of Treg cells, and resulted in alterations of the gut microbiota, including changes in beta diversity, the Firmicutes/Bacteroidetes (F/B) ratio, and the abundance of Clostridia in EAMG mice. We further conducted FMT tests and demonstrated that the EAMG Abx-treated mice which were transplanted the feces of mice treated with AS-IV significantly alleviated myasthenia symptoms, reduced Th1 and Th17 cells levels, and increased Treg cell levels.This study speculated that AS-IV ameliorates EAMG by regulating CD4 + T cells and altering the structure and species of gut microbiota of EAMG.
Abstract Background Myasthenia gravis (MG) is an acquired autoimmune disease with high heterogeneity. The disease is chronic, relapsing repeatedly and progressive with acute exacerbation occasionally. Although the treatment of MG has developed, it is still unsatisfactory and has some unexpected side effects. Traditional Chinese medicine (TCM) has shown great potential in MG treatment, including relief of muscle weakness syndrome, improvement of patient’s quality of life, and reduction of side effects of western medicine. The purpose of this study is to evaluate the effectiveness of modified Buzhong Yiqi decoction (MBYD) as an add-on therapy for MG through a small series of N-of-1 trials. Methods Single-centre, randomized, double-blind, 3 crossover N-of-1 trials will be conducted to enroll patients with MG diagnosed as spleen-stomach deficiency syndrome or spleen-kidney deficiency syndrome in TCM. Each N-of-1 trial has 3 cycles of two 4-week periods containing the MBYD period and placebo period. The wash-out interval of 1 week is prior to switching each period. Primary outcome: quantitative myasthenia gravis (QMG). Secondary outcomes: the following scales: myasthenia gravis composite (MGC), myasthenia gravis activities of daily living profile (MG-ADL), myasthenia gravis quality of life (MG-QOL); the level of CD4+FoxP3+Treg cells and cytokines (IL-4, IL-17A, INF-γ, TGF-β) in the peripheral blood; the alterations of the composition of gut microbiota; reduction of the side effects of western medicine. Discussion Used by WinBUGS software, we will conduct a hierarchical Bayesian statistical method to analyze the efficacy of MBYD in treating MG in individuals and populations. Some confounding variables such as TCM syndrome type and potential carryover effect of TCM will be introduced into the hierarchical Bayesian statistical method to improve the sensitivity and applicability of the trials, and the use of prior available information within the analysis may improve the sensitivity of the results of a series of N-of-1 trials, from both the individual and population level to study the efficacy of TCM syndrome differentiation. We assumed that this study would reveal that MBYD is effective for MG and provide robust evidence of the efficacy of TCM to treat MG. Trial registration Chinese Clinical Trial Register, ID: ChiCTR2000040477 , registration on 29 November 2020.
Introduction: The quantitative myasthenia gravis score is a commonly used scale for evaluating muscle weakness associated with myasthenia gravis (MG). It has been reported that some items used in the scale have low discriminative properties. However, there has been no research investigating the applicability of the quantitative MG score (QMGS) in Chinese patients with MG. In addition, the scoring method and ranges of grip strength items in QMGS need to be further evaluated. Methods: This study included 106 Chinese patients with MG, enrolled between September 2020 and February 2021, who were evaluated using the QMGS. Each item in the QMGS was analyzed for distribution. Three methods of evaluating grip strength, grip strength decrement, maximum grip strength, and relative grip strength, were compared. The correlation between the QMG total score minus grip strength score, and three evaluating methods, was analyzed. Results: The grip strength, swallowing, speech, diplopia, ptosis, and facial muscles items showed a clustered distribution. Most patients (94%) presented their maximum grip strength in the first four grip strength measurements. The QMG total score minus the grip strength score had a weak correlation with grip strength decrement (R grip r = 0.276; L grip r = 0.353, both p < 0.05) and moderate correlations with maximum grip strength (R grip r = -0.508; L grip r = -0.507; both p < 0.001) and relative grip strength (R grip r = -0.494; L grip r = -0.497, both p < 0.001). Conclusions: This study suggested that partial items in the QMGS have low discriminative properties for Chinese populations and the maximum grip strength value is the better method to evaluate grip strength compared to the other two scoring methods. Based on the quartiles of maximum grip strength, we propose new scoring ranges for the grip strength items.
Abstract Background : Myasthenia gravis (MG) is an acquired autoimmune disease with high heterogeneity. The disease is chronic, relapsing repeatedly and progressive with acute exacerbation occasionally. Although the treatment of MG has developed, it is still unsatisfactory and has some unexpected side effects. Traditional Chinese medicine(TCM) has shown great potential in MG treatment, including relief of muscle weakness syndrome, improvement of patient’s quality of life and reduction of side effects of western medicine. The purpose of this study is to evaluate the effectiveness of Modified Buzhong Yiqi Decoction (MBYD) as an add-on therapy for MG through a small series of N-of-1 trials. Methods : Single-centre, randomized, double-blind, 3 crossover N-of-1 trials will be conducted to enroll 12 adult patients with MG diagnosed as spleen-stomach deficiency syndrome or spleen-kidney deficiency syndrome in TCM. Each N-of-1 trial has 3 cycles of two 4-week periods containing the MBYD period and placebo period. The wash-out interval of 1-week is prior to switching each period. Primary outcome: Quantitative Myasthenia Gravis (QMG). Secondary outcomes: the following scales: myasthenia gravis composite (MGC), myasthenia gravis activities of daily living profile (MG-ADL), myasthenia gravis quality of life (MG-QOL); the level of CD4+FoxP3+Treg cells and cytokines (IL-4, IL-17A, INF-γ, TGF-β) in the peripheral blood; the alterations of the composition of gut microbiota; reduction of the side effects of western medicine. Discussion : Paired t tests or Wilcoxon signed rank tests will be used to analyse this trials’ data and the results we arrived at will be interpreted cautiously. We assumed that this study would reveal that MBYD is effective for MG and provide robust evidence of the efficacy of TCM to treat MG. Trial registration : Chinese Clinical Trial Register, ID: ChiCTR2000040477, registration on 29 November 2020, http://www.chictr.org.cn/showprojen.aspx?proj=64688.
Myasthenia gravis (MG) is an autoimmune disease in which antibodies directly target components of the neuromuscular junction, causing neuromuscular conduction damage that leads to muscle weakness. The current pharmaceutical treatment for MG is still not ideal to address the problems of disease progression, high recurrence rate, and drug side effects. Clinical observations suggest that traditional Chinese medicine (TCM) can strengthen immunity and improve symptoms of MG patients, delay the progression of the disease, reduce or even prevent the need for immunosuppressive therapy when used in combination with acetylcholinesterase inhibitors or low-dose prednisone, as well as improve the quality of life of patients. The Qiangji Jianli Capsule (QJC) is a combination of medicinal herbs which is used in traditional Chinese medicine. Since MG is a rare disorder, randomized controlled trials comparing large cohorts are difficult to conduct. Therefore, we proposed to aggregate data from a small series of N-of-1 trials to assess the effect of the Chinese medical prescription QJC, which strengthens the spleen and nourishes Qi, as an add-on treatment for MG with spleen and stomach Qi deficiency syndrome.Single-center, randomized, double-blind, multiple crossover N-of-1 studies will compare QJC versus placebo in 5 adult MG patients with spleen and stomach Qi deficiency syndrome. Patients will undergo 3 cycles of two 4-week intervention periods. According to the treatment schedule, patients will continue to be treated with pyridine bromide tablets, prednisone acetate, tablets and/or tacrolimus capsules throughout the entire trial. Each period consisting of 4-week oral add-on treatment with QJC will be compared with 4-week add-on treatment with a placebo. The primary endpoints are quantitative myasthenia gravis (QMG) test; measurement of the amount of Treg cells and cytokines such as interferon-γ (IFN-γ), interleukin-4 (IL-4), interleukin-17A (IL-17A), and transforming growth factor-β (TGF-β); and corticosteroid or immunosuppressive agent dosage. Secondary outcome measures: Clinical: Evaluation of the effect of TCM syndromes; MG-activities of daily living (MG-ADL) scales; adverse events.This study was approved by The First Affiliated Hospital of Guangzhou University of Chinese Medicine (GZUCM), No. ZYYECK[2019]038. The results will be published in a peer-reviewed publication. Regulatory stakeholders will comment on the suitability of the trial for market authorization and reimbursement purposes. Trial registration: Chinese Clinical Trial Register, ID: ChiCTR2000033516. Registered on 3 June 2020, http://www.chictr.org.cn/showprojen.aspx?proj=54618.
Ethnopharmacological relevance: Scutellaria baicalensis Georgi (SBG) is a traditional Chinese medicine widely used for its anti-inflammatory and antioxidant properties. Wogonin is one of the primary bioactive components of it. Acetaminophen(APAP)-induced liver injury(AILI) represents one of the most prevalent forms of drug-induced liver damage, primarily driven by inflammatory responses and oxidative stress.Aim of study: To investigate the specific effects and underlying mechanisms of Wogonin on AILI.Materials and methods: C57BL/6 J mice were pre-treated with Wogonin (1, 2.5, 5 mg/kg) for 3 days before APAP (300 mg/kg) administration. Serum and liver tissues were collected 24 hours post-APAP treatment. Bone marrow-derived macrophages (BMDMs) and RAW264.7 were cultured and pre-treated with Wogonin (5, 10, 20 μM) for 30 minutes, followed by stimulation with lipopolysaccharide (LPS, 100 ng/ml) for three hours before collection. To ascertain the involvement of the PI3K/AKT signaling pathway in Wogonin's protective effect against AILI, LY294002 (a PI3K inhibitor) and MK2206 (an AKT inhibitor) were used in mice and cells.Results: In mice, Wogonin pre-treatment was found to alleviate AILI in a dose-dependent manner, concurrently reducing oxidative stress and inflammatory responses. Additionally, liver transcriptome analysis indicated that Wogonin primarily affects immune function and cytokine regulation in AILI. Subsequent findings revealed that Wogonin diminishes inflammatory responses by inhibiting the PI3K/AKT signaling pathway in macrophages. Moreover, Wogonin was observed to be protective against AILI via the same pathway in mice.Conclusions: This study demonstrates that Wogonin effectively ameliorates AILI and modulates APAP-induced hepatotoxicity through the PI3K/AKT signaling pathway in mice.
Scutellaria baicalensis Georgi (SBG), a widely used traditional Chinese medicine, exhibits anti-inflammatory and antioxidant properties. Wogonin is one of the primary bioactive components of SBG. Acetaminophen (APAP)-induced liver injury (AILI) represents a prevalent form of drug-induced liver damage and is primarily driven by inflammatory responses and oxidative stress.
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